Forms of cidofovir

ABSTRACT

Cidofovir is obtained in different forms, including amorphous cidofovir, crystalline anhydrous cidofovir, crystalline cidofovir monohydrate, and crystalline cidofovir dihydrate, including various polymorphs.

CROSS-REFERENCE TO RELATED APPLICATION

This application claims priority from U.S. provisional application61/472,843, filed Apr. 7, 2011, and incorporated herein by reference inits entirety for all purposes.

FIELD OF THE INVENTION

The present invention generally relates to amorphous and crystallineforms of cidofovir, including polymorphs of crystalline cidofovir, andprocesses for preparing amorphous and crystalline forms of cidofovir. Inparticular, the invention is related to novel processes for preparingthe amorphous form and the anhydrous, monohydrate, and dihydratecrystalline forms of cidofovir, including various polymorphs.

BACKGROUND OF THE INVENTION

Solids exist in either amorphous or crystalline forms. An anhydrouscompound is a compound that does not contain water, especially water ofcrystallization. A hydrate is any compound containing water in the formof H₂O molecules, usually, but not always, with a definite content ofwater by weight. The best-known hydrates are crystalline solids thatlose their fundamental structures upon removal of the bound water. Inthe case of crystalline forms, molecules are positioned inthree-dimensional lattice sites. When a compound recrystallizes from asolution or slurry, it may recrystallize with different latticearrangements, and the different crystalline forms are sometimes referredto as “polymorphs.” The different crystalline forms or polymorphs of agiven substance may differ from each other with respect to one or morephysical properties (e.g., mechanical strength, compaction behavior,flow properties, particle size, shape, melting point, degree ofhydration or salvation, caking tendency, compactability withexcipients), chemical properties (e.g., dissolution rate, solubility),and/or biological properties (e.g., bioavailability, pharmacokinetics).The variation in properties among different crystalline forms oftenmakes one crystalline form more desirable or preferred over other forms.

Cidofovir's chemical formula is C₈H₁₄N₃O₆P and its IUPAC name is({[(S)-1-(4-amino-2-oxo-1,2-dihydropyrimidin-1-yl)-3-hydroxypropan-2-yl]oxy}methyl)phosphonic acid. Cidofovir has alsobeen described as(S)-(1-(4-amino-2-oxopyrimidin-1(2H)-yl)-3-hydroxypropan-2-yloxy)methylphosphonic acid as well as possibly by other chemical names. Itschemical structure is:

Cidofovir was discovered at the Institute of Organic Chemistry andBiochemistry, Prague, and developed by Gilead Sciences. Today, cidofoviris an injectable antiviral medication for the treatment ofcytomegalovirus (CMV) retinitis in patients with AIDS. It suppresses CMVreplication by selective inhibition of viral DNA polymerase andtherefore prevention of viral replication and transcription. It is anacyclic nucleoside phosphonate, and is therefore independent ofphosphorylation by viral enzyme, in contrast to, for instance,acyclovir.

Cidofovir is marketed with the brand name Vistide by Gilead in theUnited States and by Pfizer in other parts of the world. Vistide® is asterile, hypertonic aqueous solution for intravenous infusion only. Thesolution is clear and colorless. It is supplied in clear glass vials,each containing 375 mg of anhydrous cidofovir in 5 mL aqueous solutionat a concentration of 75 mg/mL. The formulation is pH-adjusted to 7.4with sodium hydroxide and/or hydrochloric acid and contains nopreservatives. Renal impairment is the major toxicity of Vistide®.

Presently, there are no Orange Book patents listed as having claimswhich cover Vistide , although previously U.S. Pat. No. 5,142,051 waslisted in the Orange Book for Vistide®. The '051 patent is not directedspecifically to cidofovir or its crystalline forms. Instead, it broadlydiscloses N-phosphonylmethoxyalkyl derivatives of pyrimidine and purinebases.

Because cidofovir has been demonstrated as an effective treatment forpatients with AIDS, improved forms of the compound are desired.

SUMMARY OF THE INVENTION

The invention provides crystalline forms of cidofovir and processes forproducing crystalline forms of cidofovir. Among the various aspects ofthe invention is a provision for an amorphous form of cidofovir as wellas two anhydrous crystalline forms of cidofovir (Forms I and II).

Another aspect of the invention is a provision for a monohydratecrystalline form of cidofovir (Form III).

Still another aspect of the invention is a provision for two dihydratecrystalline forms of cidofovir (Forms IV and V).

Yet another aspect of the invention is a provision for mixtures ofvarious forms of cidofovir.

Another aspect of the invention provides an amorphous (non-crystalline)form of cidofovir.

A further aspect of the invention encompasses pharmaceuticalcompositions comprising, or prepared from, amorphous cidofovir and/orcrystalline cidofovir Form I and/or crystalline cidofovir Form II and/orcrystalline cidofovir Form III and/or crystalline cidofovir Form IVand/or crystalline cidofovir Form V.

An additional aspect of the invention provides processes for preparingamorphous cidofovir and polymorphs of anhydrous, monohydrate anddihydrate forms of cidofovir.

The present invention further provides processes for obtainingsubstantially pure amorphous and crystalline forms of cidofovir or, ifdesired, mixtures of different forms of cidofovir.

A pharmaceutical formulation is further provided by the invention, whichis prepared by combining at least one form of cidofovir selected fromthe group consisting of amorphous cidofovir, cidofovir Form I, cidofovirForm II, cidofovir Form III, cidofovir Form IV and cidofovir Form V withat least one pharmaceutically acceptable excipient such as water.

Also provided by the invention is a method of treating a disease,comprising administering to a patient in need of treatment atherapeutically effective amount of at least one form of cidofovirselected from the group consisting of amorphous cidofovir, cidofovirForm I, cidofovir Form II, cidofovir Form III, cidofovir Form IV andcidofovir

Form V. The invention also provides a method of treating a disease,comprising administering to a patient in need of treatment atherapeutically effective amount of a pharmaceutical formulationprepared by combining at least one form of cidofovir selected from thegroup consisting of amorphous cidofovir, cidofovir Form I, cidofovirForm II, cidofovir Form III, cidofovir Form IV and cidofovir Form V withat least one pharmaceutically acceptable excipient such as water.

Other aspects, features and objectives of the invention will be in partapparent and in part described in more detail below.

DESCRIPTION OF THE FIGURES

FIG. 1 represents an X-ray powder diffraction (XRPD) pattern of ananhydrous crystalline form of cidofovir (Form I).

FIG. 2 represents an X-ray powder diffraction (XRPD) pattern of anothersample of cidofovir Form I.

FIG. 3 represents an XRPD pattern of another anhydrous crystalline formof cidofovir (Form II).

FIG. 4 represents an XRPD pattern of a monohydrate crystalline form ofcidofovir (Form III).

FIG. 5 represents an XRPD pattern of a dihydrate crystalline form ofcidofovir (Form IV).

FIG. 6 represents an XRPD pattern of another dihydrate crystalline formof cidofovir (Form V).

FIG. 7 shows a flow diagram outlining various aspects of the invention,including the synthesis of cidofovir.

DETAILED DESCRIPTION OF THE INVENTION

Polymorphism is defined as “the ability of a compound to crystallize inmore than one distinct crystal species” and different crystalarrangements of the same chemical composition are termed polymorphs.Polymorphs of the same compound arise due to differences in the internalarrangement of atoms and have different free energies and thereforedifferent physical properties such as solubility, chemical stability,melting point, density, flow properties, bioavailability and so forth.

It has been discovered that cidofovir can be prepared in well-definedand consistently reproducible crystalline forms, as well as mixtures ofthese forms. More specifically, the inventors have surprisingly foundthat changing the solvent composition and/or the pH used during productisolation results in different hydrated and polymorphic forms anddifferent impurity profiles being generated. Also, reliable and scalablemethods for converting hydrate forms have been developed. Moreover, thecidofovir polymorphs provided by the present invention meetInternational Conference on Harmonisation (ICH) guidelines and areuseful active ingredients in pharmaceutical formulations.

One aspect of the invention provides cidofovir Form I. Cidofovir Form Imay be characterized by an XRPD pattern substantially in accordance withthat shown in FIG. 1, an XRPD 2-theta/intensity/d-value patternsubstantially in accordance with that shown in Table A, and/or an XRPDpattern having main peaks expressed as 2-theta at about 15.8, 13.5,24.9, 21.3, 27.0, 18.6, 25.2, and 23.8 degrees (with “about” hereinbeing understood as modifying each of the stated values). Cidofovir FormI may be in admixture with one or more other forms of cidofovir or maybe substantially free of any other physical forms of cidofovir.

Another aspect of the invention provides cidofovir Form IL CidofovirForm II may be characterized by an XRPD pattern substantially inaccordance with that shown in FIG. 3, an XRPD 2-theta/intensity/d-valuepattern substantially in accordance with that shown in Table C, and/oran XRPD pattern having main peaks expressed as 2-theta at about 11.5,19.0, 18.4, 29.7, 26.2, 17.6, 17.4, 26.6, 23.1 and 21.3 degrees.Cidofovir Form II may be in admixture with one or more other forms ofcidofovir or may be substantially free of any other physical forms ofcidofovir.

In still another aspect of the invention cidofovir Form III is provided.Cidofovir Form III may be characterized by an XRPD pattern substantiallyin accordance with that shown in FIG. 4, an XRPD2-theta/intensity/d-value pattern substantially in accordance with thatshown in Table D, and/or an XRPD pattern having main peaks expressed as2-theta at about 5.5, 18.3, 14.2, 26.9, 21.1, 15.2, 19.9, 27.5, 21.3,22.0, 7.6 and 22.3 degrees. Cidofovir Form III may be in admixture withone or more other forms of cidofovir or may be substantially free of anyother physical forms of cidofovir.

Cidofovir Form IV is provided in yet another aspect of the invention.Cidofovir Form IV may be characterized by an XRPD pattern substantiallyin accordance with that shown in FIG. 5, an XRPD2-theta/intensity/d-value pattern substantially in accordance with thatshown in Table E, and/or an XRPD pattern having main peaks expressed as2-theta at about 11.5, 18.4, 19.0, 29.7, 17.6, 26.6, 26.2 and 17.4degrees. Cidofovir Form IV may be in admixture with one or more otherforms of cidofovir or may be substantially free of any other physicalforms of cidofovir.

A further aspect of the invention provides cidofovir Form V, which maybe characterized by an XRPD pattern substantially in accordance withthat shown in FIG. 6, an XRPD 2-theta/intensity/d-value patternsubstantially in accordance with that shown in Table F, and/or an XRPDpattern having main peaks expressed as 2-theta at about 17.6, 29.7,19.0, 21.3, 18.4, 26.5, 26.2 and 11.5 degrees. Cidofovir Form V may bein admixture with one or more other forms of cidofovir or may besubstantially free of any other physical forms of cidofovir.

A method of preparing anhydrous cidofovir is provided by the presentinvention, comprising isolating a composition comprised of cidofovirmonohydrate at a pH of about 2.5 to about 5.5, combining the compositionwith water to form a mixture, heating the mixture at a temperature offrom about 50° C. to about 80° C., cooling the mixture below about 50°C., and combining the mixture with ethanol to form a slurry comprisinganhydrous cidofovir.

An additional aspect of the invention provides amorphous cidofovir,which is non-crystalline and which may be characterized by thesubstantial absence of any peaks in its XRPD pattern. The amorphouscidofovir may be in admixture with one or more other forms of cidofoviror may be substantially free of any other physical form of cidofovir.

A yet further aspect of the invention provides a method of makingcidofovir dihydrate, comprising treating a composition comprised ofcidofovir monohydrate isolated at a pH of about 4.5 to about 5.5 withwater at a temperature of from about 50° C. to about 80° C. andcombining with a volume of ethanol effective to precipitate cidofovirdihydrate. Another method of making cidofovir dihydrate is also providedby the invention, comprising combining a composition comprisingcidofovir monohydrate isolated at a pH of about 4.5 to about 5.5 withwater to form a mixture, acidifying the mixture, and combining themixture with ethanol. Still another method of making cidofovir dihydrateprovided by the invention comprises slurrying a composition comprisingcidofovir monohydrate with aqueous ethanol at a temperature of fromabout 15° C. to about 35° C.

A method of making cidofovir Form III is further provided by theinvention, comprising treating a solution of cidofovir with a base toachieve a pH of about 4.5 to about 5.5 and combining the solution withan amount of ethanol effective to cause precipitation of cidofovir.

FIG. 7 is a flow diagram outlining various aspects of the invention,which are explained in further detail below.

As indicated by FIG. 7, a first step in making cidofovir involves acytosine coupling reaction in which compound 1 is coupled to compound 2to provide compound 3. The coupling reaction may be promoted by use of abase. A benzoylation step provides compound 4. De-tritylation andalkylation converts compound 4 to compound 5. In this step, compound 4is reacted with ClCH₂P(O)Cl₂ to replace the trityl group with—P(O)(Cl)CH₂Cl.

Compound 5 is hydrolyzed to compound 6 by, for example, stirring intetrahydrofuran (THF), acetonitrile (ACN), water and NEt₃(triethylamine). It is also preferred that the stirring takes place fora minimum of 1 hour. This reaction mixture is filtered and the solidsare washed with THF. If desired, the filtrate is concentrated and washedor chased with solvent to minimize the amount of water in the filtrate.

Preferably, the filtrate is chased with the solvent more than once. Itis also preferred that the solvent is a toluene/methanol mixture, morepreferably a 1:1 toluene/methanol mixture. It is further preferred thatthe amount of water present in the filtrate is reduced to a Karl Fischerresult of less than 1.0%.

In the next step, compound 6 is converted to compound 7, whereindeprotection (removal of the Bz group) and migration of the—P(O)(Cl)CH₂Cl group to the deprotected site take place. For example,the concentrated filtrate material containing compound 6 may be combinedwith an alcohol, e.g., methanol. In one embodiment, the amount ofalcohol added is 3 to 4 volumes based on compound 5. NaOCH₃ (e.g., 4.9eq) is also charged. In one embodiment, this reaction mixture is heatedto 30-35° C. and refluxed until the deprotected compound 6 startingmaterial is <1.0% by HPLC. Refluxing may, in one embodiment, be carriedout for a minimum of 20 hours.

Next, the solids are filtered and washed with methanol and the combinedfiltrate is treated with a cation exchange resin to an acidic pH.Preferably, the resin is Dowex® 50WX8 100-200 (H) resin and the pH is3.0 to 3.5. After filtration and washing of the resin with water, thefiltrate (containing compound 7) is concentrated. Preferably, thefiltrate is concentrated to 3 mL/g of the calculated initial amount ofcompound 5.

Compound 7 is then hydrolyzed to cidofovir (compound 8). In this step,concentrated hydrochloric acid (8.4 eq) may be charged to theconcentrated filtrate and the mixture heated until the compound 7starting material is less than about 1.0% by HPLC. Preferably, themixture is heated to a temperature of about 85 to 90° C. for at least 20hours. This solution is cooled and filtered through a micron filter,preferably a 0.2 micron filter. Ammonium hydroxide, preferably at aconcentration of about 30%, is charged to the solution until a pH ofabout 5.0 is attained. After a constant pH is achieved, ethanol ischarged and the solution is stirred. Preferably, 1 volume of ethanol ischarged and the solution is stirred for at least 15 minutes. As aprecipitate may form during this step, it may be necessary to add moreethanol to the solution/slurry before cooling. After mixing, thesolution or mixture is cooled, preferably, to about 5±5° C. and stirred,preferably for at least 12 hours.

At this point, the resultant solid (compound 8, crude cidofovir) can becollected, e.g., by filtration, washed with an alcohol/water mixture,preferably 2:1 EtOH/H₂O, and dried under vacuum. Preferably, the solidis dried to a point where there is no weight loss over time (i.e., to aconstant weight) at a temperature of about 40° C. The resultant solid iscrystalline cidofovir (as a monohydrate).

The above-described method is exemplary. Other methods of synthesizingcidofovir, including methods known in the art, may be utilized.

The relatively crude cidofovir obtained by the foregoing procedure maybe further purified and converted to the dihydrate form as follows. Thecrystalline cidofovir (monohydrate) is slurried in water, preferably toa concentration of about 3 mL/g. The pH is increased, preferably toabout pH 6-8, in order to dissolve the cidofovir. It is also preferredthat the pH is increased using ammonium hydroxide, most preferably at aconcentration of about 28-30%. If desired, activated carbon can becharged to reduce the color of the solution. Preferably, the carbon isadded for a period of time until a pale straw colored solution isachieved. More preferably, the carbon is a 12×20 mesh 30% (wt/wt) and ischarged and stirred for about 4 to 5 hours. The carbon is filteredthrough a micron filter, preferably a 0.2 micron filter, and washed withwater (1.5 weight/g crude and 1.0 weight/g crude). All solutions arepolish filtered. The pH of the combined filtrate and washes is thenreduced, preferably to about pH 5 using a concentrated hydrochloric acid(e.g., 3 M) and ethanol (6 volume) mixture. This results in a slurrywhich is then cooled, preferably to about 5±5° C., and stirred,preferably for at least 12 hours. The solid portion of the slurry isisolated by filtration, washed with an alcohol/water mixture, preferably2:1 EtOH/H₂O, and dried. Preferably, the filtered solid is dried toachieve a constant weight under vacuum at a temperature of about 40° C.

The isolated solid is then combined with water, preferably to aconcentration of 5 mL/g. Concentrated HCl, preferably 3 M HCl, is addedto obtain a pH of about pH 3.0-3.5. Alcohol, preferably 3 volumes ofethanol, is charged and the mixture is cooled and stirred. Preferably,the temperature of the mixture is reduced to about 5±5° C. after whichstirring occurs for at least 12 hours. The precipitated solid is thenisolated by filtration and washed, preferably with 2:1 EtOH/H₂O. Dryingto constant weight under vacuum at 40° C. results in cidofovir dihydrate(compound 9).

Four separate HPLC methods have been identified for monitoring theprocess, intermediates and final product, including one to confirm thechiral purity of the final product. The methods have been optimized andshown to be scientifically robust. Impurity marker qualification wasconducted resulting in more than 20 potential impurities in the processalthough a number of the impurities are only observed in trace levels.

Methods for determining residual levels of genotoxic impurities andreagents have been developed although so far it is believed that such amethod is only required for residual DMAP reagent, which is used duringthe first stage of the process and is removed during later processingstages.

An appropriate gas chromatography (GC) method for determining residualsolvent levels in the final product was developed. Stability indicatingmethods were also developed and conducted on the final products.

A full monograph for the final crystalline cidofovir dihydrate product(compound 9) has been completed and it was confirmed that, with an OELof 0.6 μgm⁻³, the product remains within category band 5 (safebridgecategory 3). This assessment also suggests that processing intermediatesup to and including compound 6 can be handled with a lower level ofcontainment than compounds 7 to 9.

The inventors have also discovered the following features of theirinvention as a result of conducting numerous experiments:

Preparation Procedures:

1. Isolations at pH 3.5-5 (see, e.g., FIG. 7) yield various hydratedforms of cidofovir, including dihydrate, depending on the solventconditions used.2. Isolations at pH 3.5 result in high levels of a residual impurity(uracil impurity) and require multiple treatments to reduce levelswithin specification. Note: this impurity can be reduced by adjustingthe pH to 5 using ammonium hydroxide and precipitating the material withethanol (3 vol).3. Isolations at pH 5 result in high quality product (within ICHguidelines) in various hydrated forms depending on the solventconditions used. Treatment of the crude cidofovir isolated at pH 5 withvarious treatments gives high purity HPLC material.

Interconversions:

4. Cidofovir dihydrate may be prepared by a process comprising treatinga composition comprised of cidofovir monohydrate isolated at a pH ofabout 4.5 to about 5.5 with water at a temperature of from about 50° C.to about 80° C. and combining with a volume of ethanol effective toprecipitate cidofovir dihydrate. For example, treatment of the crudematerial (monohydrate or mixtures) isolated at pH 5 with 6.6 mL water/gat 65° C. followed by precipitation with 1 vol ethanol gives purecidofovir dihydrate. In another suitable process for obtaining cidofovirdihydrate, a composition comprised of cidofovir monohydrate isolated ata pH of about 4.5 to about 5.5 is combined with water to form a mixture,the mixture is acidified (e.g., to a pH of about 2.5 to about 4 or about3 to about 3.5) using a suitable acid (e.g., HCl) and combined withethanol. The mixture may be cooled below room temperature (e.g., toabout 0° C. to about 10° C.) and stirred for a period of time (e.g.,about 6 to about 24 hours, preferably at least about 12 hours) toprovide pure cidofovir dihydrate. The cidofovir dihydrate may becollected by a suitable method (e.g., filtration), washed and/or dried.5. Isolation of cidofovir at a pH of about 2.5 to about 5.5 (e.g., 3,3.5 or 5) followed by different precipitation protocols can give rise toanhydrous forms of cidofovir. The crude cidofovir material (containingcidofovir monohydrate) does not dissolve in water at 65° C.-30 mL waterwas used and the mixture was cooled to 40° C. and 1 vol EtOH added togive a material with 0.2% KF (0.2% water as measured by Karl Fischer).The invention thus provides a method of making anhydrous cidofovir (FormI or Form II) comprising isolating a composition comprised of cidofovirmonohydrate at a pH of about 2.5 to about 5.5, combining the compositionwith water to form a mixture, heating the mixture at a temperature offrom about 50° C. to about 80° C., cooling the mixture below about 50°C., and combining the mixture with ethanol to form a slurry comprisinganhydrous cidofovir. The use of relatively larger amounts of ethanolfavors the formation of the Form I polymorph. For example, if the amountof ethanol is approximately 1 volume per volume of water, the Form IIpolymorph may be obtained, whereas if the amount of ethanol isapproximately 7 volumes per volume of water, the Form I polymorph may beobtained. The anhydrous cidofovir may be isolated by a suitableseparation means (e.g., filtration), washed and/or dried.6. Conversion from a form with a Karl Fischer value of about 7%(corresponding to a monohydrate) to a form with a Karl Fischer value ofabout 11.3% (corresponding to a dihydrate) is possible by slurrying thecrude material at about room temperature with aqueous ethanol (e.g., 1:1EtOH/H₂O). The invention thus provides a method of making cidofovirdihydrate, comprising slurrying a composition comprised of cidofovirmonohydrate with aqueous ethanol at a temperature of from about 15° C.to about 35° C.

Cidofovir monohydrate Form III may be obtained by a method comprisingtreating a solution of cidofovir with a base to achieve a pH of about4.5 to about 5.5 and combining the solution with an amount of ethanoleffective to cause precipitation of cidofovir. The initial cidofovirsolution may, for example, be an acidified solution obtained as a resultof hydrolyzing compound 7 in FIG. 7. Such solution, following thehydrolysis step (which typically involves treatment of compound 7 with astrong acid such as HCl and heating to an elevated temperature, e.g.,about 75° C. to about 100° C., for a period of time effective to achievehydrolysis of the phosphate ester group), may be filtered to remove anyresidual solid impurities prior to treatment with a base (e.g., ammoniumhydroxide). After combining with ethanol (which may be added inportions), the resulting mixture may be cooled (e.g., to a temperatureof from about 0° C. to about 10° C. and stirred for a period of time(e.g., at least about 12 hours) before isolating the precipitatedcidofovir monohydrate Form III by a suitable means such as filtration.The collected precipitate may be washed (e.g., with aqueous ethanol) andthen dried to constant weight (e.g., at a temperature of about 30° C. toabout 50° C. under vacuum).

Amorphous cidofovir may be obtained by freeze-drying (lyophilizing) asolution of cidofovir as well as by precipitating cidovir from anaqueous solution. For example, an aqueous solution of cidofovir may beprepared (e.g., by combining solid cidofovir and water and addingammonium hydroxide until dissolution is achieved). An acid such as HClmay then be added to lower the pH to about 3.5 to about 4.5 (e.g., a pHof about 4) and ethanol thereafter added to obtain precipitated solids.The mixture may be cooled below room temperature (e.g., −20 to −15° C.)for a period of time (e.g., 1 to 24 hours) and the solids then isolated(by filtration, for example), washed (with aqueous ethanol, forexample), and dried (e.g., under vacuum at 20 to 40° C.) to provideamorphous cidofovir.

Mixtures of the above-described different cidofovir forms may beobtained by varying the isolation and recrystallization as well as byseparately preparing different forms as pure substances and combiningsuch pure substances together.

A pharmaceutical formulation may be prepared by combining or formulatingat least one form of cidofovir selected from the group consisting ofamorphous cidofovir, cidofovir Form I, cidofovir Form II, cidofovir FormIII, cidofovir Form IV and cidofovir Form V with at least onepharmaceutically acceptable excipient. Suitable excipients include anyof the known or conventional ingredients or components useful to includein pharmaceutical formulations in addition to the active pharmaceuticalingredient(s), including, for example, carriers, diluents, solvents(e.g., water), preservatives, stabilizers, pH adjusting agents, wettingagents and the like. The pharmaceutical formulation can take the form ofpowders, suspensions, solutions, sprays, emulsions, pastes, ointmentsand the like and can be used, for example, for parenteral administration(intravenous, intradermal, intramuscular, intrethecal, etc.) as well asfor oral, rectal, intravaginal or intranasal administration or topicaladministration. According to the requirements and application form,these formulations can contain various concentrations of one or moreforms of cidofovir in accordance with the invention, from about 0.01 upto 100% by weight, for example. Pharmaceutical formulations inaccordance with the invention may be utilized to treat any of theconditions or diseases where cidofovir is known to have efficacy,including for example in the treatment of cytomegalovirus (CMV)retinitis in patients with AIDS.

In one embodiment of the invention, a quantity of one or more forms ofcidofovir in accordance with the invention is dissolved in water (e.g.,water suitable for injection) to provide a solution, with the pH of thesolution being adjusted to approximately neutral (e.g., 7.4) using abase (e.g., sodium hydroxide) or acid (e.g., HCl). In one embodiment, nopreservatives are present in the solution. The concentration ofcidofovir may be adjusted as desired; for example, the cidofovirconcentration may be about 75 mg/mL (calculated as anhydrous cidofovir).Such a solution may be supplied in clear glass vials, each containing375 mg of anhydrous cidofovir in 5 mL aqueous solution. The solution maybe utilized as an injectable antiviral medication for the treatment ofcytomegalovirus (CMV) retinitis in patients with AIDS.

The invention will be illustrated in more detail with reference to thefollowing embodiments and examples, but it should be understood that thepresent invention is not deemed to be limited thereto.

The different crystalline forms of cidofovir described herein arecharacterized, for example, by reference to the 2-theta (2-θ) values oftheir main peaks in their XRPD (X ray powder diffraction) patterns. Invarious embodiments, each such value is considered to include the rangeof ±0.4, ±0.3, ±0.2 or ±0.1 from the stated value.

EXAMPLES Example 1—Amorphous Form of Cidofovir

An amorphous form of cidofovir may be obtained by freeze-drying(lyophilizing) a solution of cidofovir.

Example 2—Anhydrous Form of Cidofovir (Form I)

Crude cidofovir (1.72 g) was isolated at pH 3 and heated to 65° C. inwater (30 mL) for 2 hours. The mixture was then cooled to 30° C. afterwhich ethanol (one volume, 30 mL) was charged. A slurry formed and wasstirred overnight. The slurry was filtered to collect the solid and thenthe solid was washed with an ethanol:water (2:1) mixture before dryingthe crystalline solid. The crystalline solid was determined to beanhydrous containing only about 0.3% by weight water (by Karl Fischer).The crystalline solid (Form I) was analyzed by XRPD which provided theresults in FIG. 1 and Table A. Accordingly, Form I is characterized byits XRPD pattern having main peaks expressed as 2-theta at about 15.8,13.5, 24.9, 21.3, 27.0, 18.6, 25.2, and 23.8 degrees. The cidofovir FormI thus made is therefore substantially free of any other physical formsof cidofovir.

Example 3—Confirmation of Anhydrous Form I of Cidofovir

Crude cidofovir (1.84 g) was isolated at pH 5 and heated to 65° C. inwater (5 mL). The slurry was then cooled to 40° C. after which ethanol(one volume, 6.6 mL) was charged followed by the addition of ethanol (6volumes). The slurry was stirred overnight. The slurry was filtered tocollect the solid and then the solid was washed with an ethanol:water(2:1) mixture before drying the crystalline solid. The crystalline solidwas again determined to be anhydrous containing only about 0.3% byweight water (by Karl Fischer). The crystalline solid was analyzed byXRPD and, as shown in FIG. 2 and Table B, is confirmed to be Form I. Inthis example, the XRPD pattern of the crystalline solid displayed mainpeaks expressed as 2-theta at about 15.8, 13.5, 24.9, 27.0, 21.4, 18.6,25.6 and 29.1 degrees.

Example 4—Another Anhydrous Form of Cidofovir (Form II)

The procedure set forth in Example 3 was repeated except that there wasno addition of 6 volumes of ethanol. The resultant crystalline solid wasdetermined to be a different anhydrous crystalline form (Form II) fromthe anhydrous form (Form I) obtained in Examples 2 and 3. Thecrystalline solid was analyzed by XRPD, which provided the results shownin FIG. 3 and Table C. The XRPD pattern of the crystalline soliddisplayed main peaks expressed as 2-theta at about 11.5, 19.0, 18.4,29.7, 26.2, 17.6, 17.4, 26.6, 23.1 and 21.3 degrees.

Example 5—Monohydrate Form of Cidofovir (Form III)

Compound 5 (FIG. 6) is hydrolyzed to compound 6 by stirring intetrahydrofuran (THF) (8.5 vol), acetonitrile (ACN) (0.5 vol), H₂O (2.0eq.) and NEt₃ (triethylamine) (2.0 eq) for a minimum of 1 hour. Thisreaction mixture is filtered and the solids washed with THF. Thefiltrate is concentrated and chased with 1:1 toluene/methanol (2×) toobtain a Karl Fischer value of <1.0%. The concentrated material is takenup in methanol (3 to 4 volumes based on the amount of compound 5),warmed to 30-35° C. and NaOCH₃/MeOH (4.9 eq) is charged. This reactionmixture is heated to reflux for a minimum of 20 hours until the amountof deprotected compound 6 starting material is <1.0% by HPLC. The solidsare filtered and washed with methanol. The combined filtrate is treatedwith Dowex® 50WX8 100-200 (H) resin to pH 3.0 to 3.5. After filtrationand washing of the resin with water, the filtrate is concentrated to avolume of 3 mL/g of the initial amount of compound 5. Concentrated HCl(8.4 eq) is charged and the mixture heated to 85 to 90° C. for a minimumof 20 hours until the amount of compound 7 starting material is <1.0% byHPLC. This solution is cooled, and filtered through a 0.2 micron filter.Ammonium hydroxide (28-30%) is charged to the solution to pH 5.0. Afterconstant pH is attained, ethanol (1 volume) is charged and stirred for aminimum of 15 minutes.

Precipitation usually occurs. Additional ethanol (2 volumes) is chargedand the mixture cooled to 5±5° C. and stirred for a minimum of 12 hours.The solid is collected by filtration, washed with 2:1 EtOH/H₂O and driedto constant weight under vacuum at 40° C. to give cidofovir (as amonohydrate). The crystalline solid was determined to be a monohydratecontaining about 7% by weight water (by Karl Fischer). The crystallinesolid (Form III) was analyzed by XRPD which provided the results in FIG.4 and Table D. Accordingly, Form III is characterized by an XRPD patternhaving main peaks expressed as 2-theta at about 5.5, 18.3, 14.2, 26.9,21.1, 15.2, 19.9, 27.5, 21.3, 22.0, 7.6 and 22.3 degrees. The cidofovirForm III is substantially free of any other physical forms of cidofovir.

Example 6—Dihydrate Forms of Cidofovir (Forms IV and V)

The procedure set forth above in Example 5 was used to make themonohydrate form of cidofovir. The solid thus obtained is slurried inwater (3 mL/g). The pH is adjusted to pH 6-8 for dissolution usingammonium hydroxide (28-30%). Activated carbon 12×20 mesh 30%weight/weight is charged and stirred for 4 to 5 hours and the color ofthe solution monitored (pale straw colored solution is expected). Afterthe required stir time, the carbon is filtered off through a 0.2 micronfilter. The carbon is washed with water (1.5 weight/g crude and 1.0weight/g crude). All solutions are polish filtered. The combinedfiltrate and washes are adjusted to pH 5 with 3M HCl and ethanol (6volumes) is charged. This slurry is cooled to 5±5° C. and stirred for aminimum of 12 h. The solid is isolated by filtration, washed with 2:1EtOH/H₂O and dried to constant weight under vacuum at 40° C. to give thepH 5 solid. This isolated solid is taken up in water (5 mL/g of the pH 5solid) and 3M HCl is carefully added to a constant pH of pH 3.0 to 3.5.Ethanol (3 volumes) is charged. This mixture is cooled to 5±5° C. andstirred for a minimum of 12 hours. The solid is isolated by filtrationand washed with 2:1 EtOH/H₂O, then dried to constant weight under vacuumat 40° C. to provide cidofovir dihydrate.

The crystalline form of cidofovir dihydrate has been found to varysomewhat depending upon the conditions used to prepare it. At present,it is believed that cidofovir dihydrate has been isolated in twodifferent crystalline forms, referred to herein as Form IV and Form V,which have somewhat similar (yet different) X-ray powder diffractionpatterns. One representative sample of cidofovir dihydrate Form IV(Sample 6-2 in the table below) was analyzed by XRPD to provide theresults shown in FIG. 5 and Table E. This sample was characterized by anXRPD pattern having main peaks expressed as 2-theta at about 11.5, 18.4,19.0, 29.7, 17.6, 26.6, 26.2 and 17.4 degrees. A representative sampleof cidofovir dihydrate Form V (Sample 6-1 in the table below) wasanalyzed by XRPD to provide the results shown in FIG. 6 and Table F.This sample was characterized by an XRPD pattern having main peaksexpressed as 2-theta at about 17.6, 29.7, 19.0, 21.3, 18.4, 26.5, 26.2and 11.5 degrees.

The following table provides a summary of certain experiments conductedby the inventors for the purpose of identifying the conditions whichproduce either the Form IV or Form V of cidofovir dihydrate.

Treatment of Cidofovir Isolation of Sample No. solid solid KF Form 6-11.02 g/6.6 1 vol EtOH 11.6% V mL H₂O at at 40° C. 65° C. 6-2 Dried in pH3.5 12.1% IV oven 35° C. 6-3 Dried at pH 3.5 11.6% IV room temperature6-4 Heated in 1 vol EtOH 11.5% IV 11.5 mL at 40° C. water (5 then 6 volmL/g crude) EtOH at at 65° C. room temperature

Example 7—Amorphous Cidofovir

Intermediate 5 (FIG. 7; 0.5 g, 0.054 mol) was heated with a solution ofsodium methoxide in methanol (0.5 M, 15 mL, 7.5 mmol) at 72° C. for 14.5h then at 90° C. for 5.5 h. The reaction mixture was quenched with water(10 mL) and filtered through a bed of ion exchange resin Dowex® 50WX8100-200 (H). The filtrate was cycled through the ion exchange bed (2times) then washed successively with 1:1 methanol:water (40 mL),methanol (40 mL) and 4% triethylamine:methanol (50 mL). Thision-exchange bed was further washed with 48:48:4methanol:water:triethylamine (100 mL) until no UV absorbance wasdetected in the filtrate. This reaction produced intermediate 7 (FIG. 7)together with cyclic cidofovir impurity. This mixture was then dissolvedin 6 N HCl and heated to 65° C. After cooling the reaction mixture toroom temperature, ethyl acetate was charged and stirred and the aqueouslayer separated. The aqueous was stirred with ethanol (50 mL). Theprecipitated material was filtered and the solid was washed withethanol. The ethanol filtrate was concentrated. The concentratedmaterial was taken up in acetonitrile and stirred with trimethylsilylbromide (19 mL) at room temperature for 18 h. The reaction mixture wasfiltered and the filtrate concentrated. The residue was taken up intoluene (30 mL) and ammonium hydroxide (28%, 50 mL) was charged andstirred at room temperature. The organic phase was separated and theaqueous phase was concentrated to dryness. Water (20 mL) and ethanol (15mL) were added to the residue. The mixture pH was 6 and was adjusted topH 3 with concentrated HCl (2 mL) then adjusted to pH 4 to 4.5 with 28%NH₄OH. After stirring for 0.5 h, the mixture was cooled, filtered andthe solids washed with 2:1 EtOH:H₂O and dried under vacuum for 18 h. Theisolated solid was taken up in water (10 mL) and 28% NH₄OH added to givea solution. Concentrated HCl was added to the solution until pH 4 wasreached. Ethanol (13 mL) was charged and the mixture stirred at −17° C.for 18 h, filtered and the solids washed with 2:1 EtOH:water (2×8 mL),dried under vacuum at 35° C. The cidofovir isolated in this manner wasdetermined to be in the amorphous form by XRPD.

TABLE A Intensity % Angle Intensity d value % 2-Theta ° Count Angstrom100.0 15.828 99902 5.59455 78.8 13.525 78759 6.54140 49.3 24.870 492663.57725 32.5 21.330 32513 4.16222 30.6 26.982 30556 3.30183 23.0 18.59922984 4.76673 20.0 25.191 19940 3.53236 18.5 23.790 18454 3.73709 18.216.677 18193 5.31174 15.7 25.611 15698 3.47536 14.6 29.064 14563 3.0699212.5 19.428 12491 4.56534 10.4 28.733 10407 3.10448 10.3 32.891 103002.72089 9.5 30.183 9484 2.95857 9.0 28.539 9022 3.12520 8.4 9.388 84019.41283 6.5 33.504 6447 2.67251 6.2 41.178 6241 2.19047 5.6 36.573 56442.45499 5.2 27.311 5193 3.26279 4.8 47.197 4784 1.92419 4.2 37.555 41732.39304 4.0 40.335 4015 2.23428 4.0 10.751 4028 8.22279 3.9 38.716 39242.32388 3.8 18.974 3825 4.67349 3.8 46.477 3844 1.95232 3.8 41.388 37792.17981 3.8 35.895 3825 2.49982 3.6 30.928 3571 2.88901 3.4 49.346 34081.84532 3.4 32.374 3414 2.76314 3.2 43.575 3207 2.07536 3.2 35.178 32012.54905 3.2 40.542 3179 2.22333 3.1 17.155 3058 5.16476 3.1 44.016 31132.05560 3.1 37.201 3099 2.41499 3.1 37.748 3067 2.38124 3.0 31.354 30282.85070 2.9 40.090 2898 2.24738 2.9 12.206 2918 7.24542 2.8 42.792 28382.11151 2.7 42.023 2718 2.14837 2.7 26.043 2683 3.41871 2.6 14.254 25716.20867 2.5 32.093 2459 2.78674 2.5 35.501 2529 2.52663 2.4 51.096 24081.78613 2.4 23.313 2425 3.81253 2.4 34.796 2424 2.57621 2.3 48.103 22601.89005 2.2 53.162 2199 1.72150 2.1 51.443 2128 1.77489 2.1 27.848 21403.20110 2.1 44.919 2118 2.01632 2.0 20.005 2030 4.43490 2.0 49.833 20281.82840 1.9 52.133 1908 1.75300 1.9 22.849 1879 3.88898 1.9 39.555 18902.27653 1.8 51.738 1821 1.76547 1.7 44.625 1703 2.02893 1.6 54.507 16061.68214 1.5 48.964 1470 1.85878 1.5 46.052 1538 1.96932 1.5 52.821 14871.73178 1.4 53.687 1351 1.70589 1.3 45.403 1326 1.99597 1.2 50.441 11661.80779

TABLE B Intensity % Angle Intensity d value % 2-Theta ° Count Angstrom100.0 15.832 135966 5.59306 87.3 13.534 118705 6.53738 34.7 24.863 471483.57832 25.3 26.975 34460 3.30274 22.4 21.375 30457 4.15364 16.0 18.59621821 4.76764 15.8 25.609 21521 3.47570 13.3 29.070 18131 3.06924 12.716.675 17276 5.31230 10.7 25.195 14496 3.53188 8.9 23.789 12155 3.737307.5 28.741 10251 3.10368 7.2 28.544 9785 3.12464 7.1 32.900 9672 2.720196.3 9.378 8549 9.42343 5.9 19.445 8037 4.56143 5.1 30.196 6964 2.957344.7 36.571 6353 2.45514 4.2 19.025 5776 4.66114 4.2 27.319 5693 3.261903.9 47.183 5364 1.92472 3.9 41.184 5238 2.19015 3.9 33.506 5367 2.672403.5 38.724 4773 2.32345 3.1 35.902 4178 2.49935 2.8 41.369 3744 2.180802.8 35.186 3765 2.54854 2.6 26.052 3510 3.41759 2.6 43.735 3541 2.068112.5 40.099 3347 2.24688 2.5 40.388 3352 2.23144 2.4 30.973 3264 2.884872.4 46.494 3266 1.95162 2.3 31.349 3084 2.85113 2.3 32.381 3146 2.762612.3 37.577 3141 2.39168 2.2 10.745 3056 8.22682 2.0 37.223 2722 2.413602.0 17.153 2751 5.16522 1.9 42.751 2620 2.11344 1.9 49.339 2632 1.845561.9 17.578 2601 5.04147 1.9 44.017 2595 2.05554 1.8 35.493 2434 2.527211.8 12.212 2400 7.24197 1.8 51.088 2486 1.78639 1.8 14.231 2484 6.218761.8 11.473 2452 7.70638 1.7 34.800 2254 2.57590 1.7 27.837 2309 3.202371.7 53.168 2321 1.72129 1.7 32.076 2364 2.78818 1.6 42.017 2232 2.148661.6 23.344 2149 3.80762 1.6 48.110 2165 1.88980 1.6 44.924 2173 2.016111.5 29.657 1997 3.00981 1.5 52.126 2030 1.75322 1.5 51.429 1976 1.775341.4 49.824 1863 1.82872 1.4 22.855 1899 3.88791 1.3 39.143 1821 2.299511.3 20.024 1781 4.43073 1.2 39.617 1655 2.27307 1.2 44.627 1616 2.028841.2 48.933 1632 1.85990 1.2 46.065 1676 1.96882 1.1 54.567 1496 1.680441.1 53.676 1478 1.70620 0.9 50.485 1194 1.80632

TABLE C Intensity % Angle Intensity d value % 2-Theta ° Count Angstrom100.0 11.497 51807 7.69078 92.8 18.989 48051 4.66972 91.9 18.373 476014.82506 57.8 29.661 29967 3.00944 52.6 26.195 27260 3.39929 43.9 17.61722761 5.03016 41.3 17.416 21402 5.08800 39.2 26.600 20320 3.34840 26.923.054 13921 3.85474 20.8 21.315 10787 4.16519 18.5 27.656 9603 3.2228818.0 35.058 9305 2.55752 16.6 25.037 8587 3.55375 15.5 30.658 80202.91384 14.7 9.144 7591 9.66314 14.3 47.342 7386 1.91864 13.5 15.6947016 5.64216 13.3 39.177 6894 2.29760 13.1 40.815 6798 2.20911 12.833.025 6632 2.71021 11.7 30.219 6047 2.95516 11.2 22.615 5815 3.9285510.7 25.478 5548 3.49328 10.6 37.656 5501 2.38681 9.8 32.502 51022.75260 9.3 43.613 4841 2.07363 9.0 21.932 4642 4.04943 7.6 33.447 39202.67692 7.5 50.824 3861 1.79505 7.3 31.744 3757 2.81657 7.2 38.539 37122.33414 7.1 8.766 3704 10.07914 6.4 38.151 3325 2.35699 5.9 39.614 30592.27325 5.7 40.003 2928 2.25202 5.5 28.708 2844 3.10714 5.5 42.252 28382.13725 5.2 46.505 2684 1.95118 5.1 32.179 2622 2.77947 5.1 49.137 26531.85265 4.9 41.496 2532 2.17442 4.8 35.720 2462 2.51166 4.7 53.904 24241.69952 4.6 50.322 2387 1.81176 4.6 44.375 2390 2.03977 4.5 45.630 23571.98655 4.4 34.018 2274 2.63334 4.4 48.434 2269 1.87789 4.3 13.800 22456.41178 4.1 48.905 2115 1.86089 3.7 47.725 1910 1.90413 3.7 52.130 19411.75312 3.5 44.816 1831 2.02071 3.4 34.363 1778 2.60763 3.3 49.726 16851.83207 2.9 51.819 1480 1.76290 2.4 52.826 1223 1.73165

TABLE D Intensity % Angle Intensity d value % 2-Theta ° Count Angstrom100.0 5.538 41111 15.94646 46.1 18.281 18934 4.84898 41.8 14.181 171916.24029 36.7 26.902 15108 3.31147 31.5 21.115 12962 4.20427 30.1 15.22712380 5.81385 26.9 19.913 11041 4.45517 25.3 27.485 10419 3.24253 24.721.297 10153 4.16866 23.9 21.997 9837 4.03753 22.8 7.589 9380 11.6399220.2 22.309 8311 3.98182 19.4 20.490 7969 4.33096 17.4 19.192 71574.62084 17.3 16.837 7099 5.26138 16.8 21.789 6910 4.07564 14.6 33.8136000 2.64879 14.2 11.113 5835 7.95517 14.1 14.533 5788 6.08996 14.013.216 5767 6.69382 13.1 18.827 5386 4.70975 13.1 23.247 5377 3.8232812.4 13.764 5093 6.42847 12.3 25.246 5062 3.52478 12.2 38.672 50242.32643 12.1 26.074 4993 3.41479 12.0 32.614 4918 2.74341 11.9 26.3854903 3.37525 10.9 28.010 4487 3.18301 10.8 22.959 4448 3.87056 10.524.754 4315 3.59372 9.8 25.723 4047 3.46059 9.2 31.238 3766 2.86100 8.96.588 3670 13.40638 8.9 17.236 3658 5.14068 8.9 33.497 3671 2.67303 8.823.650 3608 3.75890 8.5 28.589 3503 3.11977 8.4 9.548 3454 9.25585 8.332.199 3429 2.77782 8.2 35.988 3365 2.49355 7.8 31.745 3198 2.81649 7.730.629 3185 2.91654 7.2 43.549 2959 2.07654 6.4 40.247 2635 2.23894 6.329.513 2577 3.02423 6.2 11.850 2542 7.46215 6.2 11.525 2557 7.67191 6.039.362 2459 2.28725 5.8 29.763 2388 2.99932 5.7 37.209 2347 2.41447 5.735.084 2341 2.55566 5.6 34.920 2288 2.56735 5.4 42.920 2230 2.10550 5.350.277 2171 1.81328 5.2 42.171 2141 2.14112 5.1 44.399 2104 2.03874 5.147.087 2105 1.92843 4.8 44.883 1984 2.01788 4.8 41.043 1964 2.19737 4.848.524 1977 1.87461 4.7 41.756 1949 2.16144 4.6 52.217 1896 1.75038 4.547.653 1833 1.90683 4.4 45.491 1797 1.99231 4.3 46.606 1753 1.94718 4.151.790 1686 1.76381 4.1 52.977 1670 1.72707 3.8 53.765 1544 1.70360 3.849.860 1566 1.82749 3.7 54.616 1505 1.67904 3.7 50.785 1540 1.79632

TABLE E Intensity % Angle Intensity d value % 2-Theta ° Count Angstrom100.0 11.509 80550 7.68288 88.9 18.375 71577 4.82446 47.7 18.996 383954.66801 39.4 29.663 31775 3.00924 37.6 17.621 30300 5.02903 31.6 26.55725471 3.35375 23.1 26.227 18586 3.39513 19.9 17.420 16012 5.08659 17.435.061 14019 2.55734 15.6 23.102 12552 3.84693 15.5 21.313 12449 4.1656313.6 47.356 10980 1.91810 11.0 9.177 8839 9.62932 10.3 30.668 82632.91285 9.8 27.656 7856 3.22295 9.1 25.062 7324 3.55037 9.0 30.229 72212.95422 8.4 40.824 6801 2.20864 8.3 39.198 6713 2.29644 8.2 33.031 65742.70969 7.1 32.540 5739 2.74944 7.0 43.619 5667 2.07334 6.8 37.675 54792.38569 6.4 15.725 5181 5.63083 6.0 22.625 4816 3.92696 5.9 50.830 47231.79485 5.8 25.490 4651 3.49160 5.1 21.952 4101 4.04565 4.8 8.775 383610.06889 4.7 38.545 3755 2.33381 4.5 5.527 3594 15.97568 4.5 33.471 36062.67508 3.9 38.157 3180 2.35664 3.9 39.611 3108 2.27342 3.8 31.760 30992.81518 3.4 40.021 2777 2.25107 3.3 49.153 2677 1.85210 3.2 35.728 25832.51109 3.2 42.280 2562 2.13589 3.0 50.348 2397 1.81091 3.0 53.901 23911.69961 3.0 45.624 2422 1.98681 3.0 28.727 2402 3.10510 2.9 46.530 22971.95020 2.8 44.398 2277 2.03878 2.7 13.814 2160 6.40525 2.7 48.945 21621.85948 2.7 41.498 2170 2.17432 2.6 10.362 2113 8.53052 2.5 52.150 20301.75249 2.5 15.984 2003 5.54035 2.4 48.440 1952 1.87768 2.4 34.048 19592.63106 2.3 14.199 1851 6.23275 2.3 13.299 1889 6.65238 2.2 19.841 17644.47120 2.1 49.823 1705 1.82874 2.1 44.825 1721 2.02033 2.1 16.568 16975.34620 2.0 34.364 1597 2.60758 1.8 51.759 1440 1.76481 1.5 52.815 12151.73198

TABLE F Intensity % Angle Intensity d value % 2-Theta ° Count Angstrom100.0 17.617 83975 5.03032 77.1 29.686 64724 3.00698 68.3 19.017 573674.66300 45.8 21.309 38434 4.16638 45.5 18.355 38226 4.82971 27.5 26.52923116 3.35722 26.8 26.247 22523 3.39266 25.7 11.491 21599 7.69436 18.925.074 15852 3.54869 9.5 23.058 7987 3.85413 9.2 30.672 7713 2.91250 8.815.712 7380 5.63553 8.4 9.137 7017 9.67130 8.0 27.742 6703 3.21314 7.821.956 6529 4.04504 7.7 8.759 6452 10.08799 7.4 25.505 6240 3.48957 5.722.608 4780 3.92982 4.6 31.785 3850 2.81301 4.0 30.194 3351 2.95753 3.38.012 2751 11.02554 3.3 13.784 2760 6.41945 3.2 28.753 2663 3.10240 2.615.984 2165 5.54035 2.1 20.667 1741 4.29422

1-17. (canceled)
 18. A method of preparing anhydrous cidofovir,comprising combining cidofovir monohydrate with water to form a mixture,heating the mixture at a temperature of from about 50° C. to about 80°C., cooling the mixture below about 50° C., and combining the mixturewith ethanol. 19-20. (canceled)
 21. A method of treating a disease,comprising administering to a patient in need of treatment atherapeutically effective amount of a pharmaceutical formulationcomprising at least one form of cidofovir, wherein the form of cidofoviris selected from the group consisting of: a) cidofovir Form I,characterized by an XRPD pattern having main peaks expressed as 2-thetaat about 15.8, 13.5, 24.9, 21.3, 27.0, 18.6, 25.2 and 23.8 degrees; b)cidofovir Form II, characterized by an XRPD pattern having main peaksexpressed as 2-theta at about 11.5, 19.0, 18.4, 29.7, 26.2, 17.6, 17.4,26.6, 23.1 and 21.3 degrees; c) cidofovir Form III, characterized by anXRPD pattern having main peaks expressed as 2-theta at about 5.5, 18.3,14.2, 26.9, 21.1, 15.2, 19.9, 27.5, 21.3, 22.0, 7.6 and 22.3 degrees; d)cidofovir Form IV, characterized by an XRPD pattern having main peaksexpressed as 2-theta at about 11.5, 18.4, 19.0, 29.7, 17.6, 26.6, 26.2and 17.4 degrees; and e) cidofovir Form V, characterized by an XRPDpattern having main peaks expressed as 2-theta at about 17.6, 29.7,19.0, 21.3, 18.4, 26.5, 26.2 and 11.5 degrees.
 22. A method of makingamorphous cidofovir, comprising freeze-drying a solution of cidofovir.23. A method of making cidofovir dihydrate, comprising combiningcidofovir monohydrate with water and adding ethanol.
 24. A method ofmaking cidofovir dihydrate, comprising combining cidofovir monohydratewith water to form a mixture, acidifying the mixture, and combining themixture with ethanol.
 25. A method of making cidofovir dihydrate,comprising combining cidofovir monohydrate with ethanol.
 26. (canceled)27. The method of claim 18, wherein the cidofovir monohydrate isisolated at a pH of about 2.5 to about 5.5.
 28. The method of claim 23,wherein the cidofovir monohydrate is isolated at a pH of about 4.5 toabout 5.5.
 29. The method of claim 23, wherein the combining occurs at atemperature of from about 50° C. to about 80° C.
 30. The method of claim24, wherein the cidofovir monohydrate is isolated at a pH of about 4.5to about 5.5.
 31. The method of claim 25, wherein the ethanol isaqueous.
 32. The method of claim 25, wherein the combining occurs at atemperature of from about 15° C. to about 35° C.